PuSH - Publication Server of Helmholtz Zentrum München: Vagaries of the ELISpot assay: Specific detection of antigen responsive cells requires purified CD8<sup>+</sup> T cells and MHC class I expressing antigen presenting cell lines.

Navigation

Home

Deutsch

Research

Advanced Search

Browse by ...

... Journal

... Publication Type

... Research Data

... Publication Year

Publication overview

Support & Contact

Contact persons

Help

Data protection

Fuchs, Y.F.* ; Jainta, G.W.* ; Kühn, D.* ; Wilhelm, C.* ; Weigelt, M.* ; Karasinsky, A.* ; Upadhyaya, B.* ; Ziegler, A.-G. ; Bonifacio, E.

Vagaries of the ELISpot assay: Specific detection of antigen responsive cells requires purified CD8⁺ T cells and MHC class I expressing antigen presenting cell lines.

Clin. Immunol. 157, 216-225 (2015)

DOI

PMC

Open Access Green as soon as Postprint is submitted to ZB.

Abstract
Metrics
Extra information

Quantification of antigen-specific CD8(+) T cells is important for monitoring infection, vaccination, and response to therapy in cancer and immune-mediated diseases. Cytokine enzyme-linked-immunospot (ELISpot) assays are often used for this purpose. We found that substantial spot formation in IFNγ ELISpot assays occurred independently of CD8(+) T cells even when classical MHC class I restricted peptides are used for stimulation. Using fractionated cells and intracellular cytokine staining, the non-CD8(+) T cell IFNγ production was attributed to the CD4(+) T cell fraction. We therefore refined a cell line-based ELISpot assay combining HLA-A*0201 expressing K562 cells for antigen presentation with purified CD8(+) T cells and demonstrated that it specifically detected CD8(+) T cell responses with detection limits comparable to traditional ELISpot assays and dextramer-based quantification. The assay was further adapted to whole antigen responses with antigen (pre-proinsulin)-expressing HLA-A*0201K562 cells. Thus, we revealed and corrected a weak spot of the CD8(+) ELISpot assay.

Altmetric

Additional Metrics?

[➜Log in]

Edit extra informations Login

Publication type Article: Journal article

Document type Scientific Article

Corresponding Author

Keywords Cd8(+) T Cells ; Elispot ; Mhc Class I ; Autoantigen ; Diabetes; Interferon-gamma; Lymphocytes-t; Nod Mice; Peptides; Binding; Identification; Molecules; Recognition; Autoantigens; Autoimmunity

ISSN (print) / ISBN 1521-6616

e-ISSN 1521-7035

Journal Clinical Immunology

Quellenangaben Volume: 157, Issue: 2, Pages: 216-225

Publisher Elsevier

Publishing Place San Diego

Non-patent literature Publications

Reviewing status Peer reviewed

Institute(s) Institute of Diabetes Research Type 1 (IDF)
Institute for Pancreatic Beta Cell Research (IPI)

PuSH - Publication Server of Helmholtz Zentrum München

Vagaries of the ELISpot assay: Specific detection of antigen responsive cells requires purified CD8+ T cells and MHC class I expressing antigen presenting cell lines.

Vagaries of the ELISpot assay: Specific detection of antigen responsive cells requires purified CD8⁺ T cells and MHC class I expressing antigen presenting cell lines.